The affinity of the Klenow fragment of E. coli DNA-polymerase 1 to primers containing bases noncomplementary to the template and hairpin-like elements

• Published in: FEBS letters Vol. 300; no. 1; pp. 18 - 20
• Main Authors: Ljach, Marija V., Kolocheva, Tatjana I., Gorn, Vladimir V., Levina, Asja S., Nevinsky, Georgy A.
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 23-03-1992
• Subjects: Abnormally high affinity; DNA Polymerase I - metabolism; Escherichia coli - enzymology; initial and maximum rates of polymerization reaction, respectively; Kinetics; Klenow fragment; Klenow fragment of E. coli DNA-polymerase 1; Nucleic Acid Conformation; Primers; Solutions; Templates, Genetic; V 0, V max; Klenow fragment; V 0, V max; Primers; FK; Abnormally high affinity
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(92)80155-A

The
K
m and
V
max values for a set of primers: d(pT)
n (pC)(pT)
m (
n=3–9.
m=0–7) and d(pT)
4 (pCpG)
4 (pT)
4 (
k=1–5) have been estimated. Poly(dA) was used as template. The number of complementary bases from the 3′ end to a noncomplementatry ones was shown to determine the efficiency of interaction of d(pT)
n (pC) (pT)
m with the Klenow fragment. Oligonucleotides d(pT)
4 (pCpG)
4 (pT)
4 in solution forming duplexes containing hairpin-like elements, show a higher affinity to the enzyme that control d(pT)
4, d(pT)
n, and d(pT)
n (pC)(pT)
m primers. For example, the
K
m value (1.1 nM) for d(pT)
4 (pCpG)
5 (pT)
4 is about 14,000 and 200 times lower than those for d(pT)
4 and d(pT)
? respectively. Possible reasons for such an abnormally high affinity of the above primers are discussed.